摘要
目的:探讨类风湿关节炎(rheumatoid arthritis,RA)滑膜成纤维样细胞(fibroblast-like synovialcells,FLS)体外培养时的增生特点,观察沙利度胺(thalidomide)、白芍总苷(total glucosides of peony,TGP)、艾拉莫德(iuratimod,T-614)在体外实验中对RA-FLS细胞增殖分化特性的影响。方法:关节镜、关节活检针取RA患者滑膜组织或抽取膝关节积液分离滑膜细胞并培养和鉴定,MTT法和反转录-聚合酶链反应(RT-PCR)法分别观察沙利度胺、TGP、T-614对FLS细胞存活分数(SF)的影响和RA-FLS的c-fos mRNA表达。结果:体外培养传代的RA-FLS为非恶性无限制增生。沙利度胺、TGP和T-614对RA-FLS的细胞染色和表面标记等特征无明显影响,但对RA-FLS的SF值均有不同程度的抑制(P<0.05);3种制剂中沙利度胺对FLS的SF抑制率最强,T-614对FLS的作用最小(P<0.05)。生理剂量的沙利度胺和高剂量TGP干预时RA组药物浓度与FLS的SF值呈负相关(P<0.05)。RA-FLS的c-fos mRNA表达率高,加入沙利度胺、TGP和T-614共孵育3 d后c-fos的表达率下降,以沙利度胺作用最明显(P<0.05)。结论:在无炎症因子刺激时,体外培养的RA-FLS呈良性增生方式。在实验剂量下沙利度胺、TGP和T-614均通过抑制c-fos的表达、降低FLS增殖能力等不同途径发挥对RA滑膜细胞的免疫调节作用。
[ABSTRACr] Objective： To investigate the proliferation of fibroblast-like synovial（FLS） cells in rheumatoid arthritis （RA） patients in vitro and the immunosuppressive effect of thalidomide, total glucosides of paeony （TGP） and T-614 on them. Methods： Synovial specimens were obtained from 6 patients with RA by arthroscopy and blindly needle biopsy, or by pumping synovial fluid on knees. FLS cells were identified in vitro by histopathological dyeing, immunohistochemical analysis and fluorescence-activated cell sorting（FCS） method for cell surface molecules respectively. The synoviocyte proliferation was measured by MTT colorimetric assay and the survival fraction（SF） of FLS was evaluated in the presence or absence of thalidomide, TGP and T-614. The expression of c-fos mRNA was examined by semi-quantitative RT-PCR in FLS cells of RA patients. Results： The identified synovial cells in patients with RA were FLS（Vimentin and Fibronectin expression positive）either cultured with agents or not. The SF of all FLS interfered by thalidomide, TGP or T-614 was lower than vacant contrast without agents in RA shown by MTF assay（ P 〈 0.05）. Thalidomide operated the strongest, whereas T-614 the weakest in the three agents（ P 〈 0.05 ）. Thalidomide at physiological doses and TGP in high concentration demonstrated marked dose-dependent inhibitory curses on FLS proliferation efficiency（ P 〈 0.05）. It was detected by semi-quantitative RT-PCR that c-fos mRNA of RA-FLS expressed highly and decreased obviously when incubation with thalidomide, TGP or T-614. In all the three agents, thalidomide down-regulated c-fos expression markedly（ P 〈 0.05 ）. Conclusion： There are no distinct activated unlimited viability in RA-FLS without being stimulated by proinflammatory cytokine in vitro. The results have confirmed our suggestion that thalidomide, TGP and T-614 possess the immunosuppressive effect to synovitis by modulating the cell proliferation and c-fos mRANA gene transcription directly i
出处
《山东大学学报（医学版）》
CAS
北大核心
2006年第11期1095-1099,共5页
Journal of Shandong University:Health Sciences
基金
山东省科技厅基金资助课题(YZ001C16)
关键词
类风湿关节炎
滑膜
成纤维细胞
沙利度胺
白芍总苷
艾拉莫德
Rheumatoid arthritis
Synovial membrane
Fibroblasts
Thalidomide
Total glucosides of paeony
T-614
